ATR mediates cisplatin resistance in 3D-cultured breast cancer cells via translesion DNA synthesis modulation

Author(s)

Gomes, Luciana Rodrigues ^{[1, 2]} ; Reily Rocha, Clarissa Ribeiro ^{[2, 3]} ; Martins, Davi Jardim ^[2] ; Petisco Fiore, Ana Paula Zen ^[4] ; Kinker, Gabriela Sarti ^[5] ; Bruni-Cardoso, Alexandre ^[4] ; Martins Menck, Carlos Frederico ^[2]

Affiliation

^[1] Inst Butantan, Lab Especial Ciclo Celular, Sao Paulo, SP - Brazil

^[2] Univ Sao Paulo, Inst Ciencias Biomed, Dept Microbiol, Sao Paulo, SP - Brazil

^[3] Univ Fed Sao Paulo, Escola Paulista Med, Dept Oncol Clin & Expt, Sao Paulo, SP - Brazil

^[4] Univ Sao Paulo, Inst Quim, Dept Bioquim, Sao Paulo, SP - Brazil

^[5] Univ Sao Paulo, Inst Biol, Dept Fisiol, Sao Paulo, SP - Brazil

• Document type: Journal article
• Source: CELL DEATH & DISEASE; v. 10, JUN 12 2019.
• Web of Science Citations: 4

Abstract

Tissue architecture and cell-extracellular matrix (cell-ECM) interaction determine the organ specificity; however, the influences of these factors on anticancer drugs preclinical studies are highly neglected. For considering such aspects, three-dimensional (3D) cell culture models are relevant tools for accurate analysis of cellular responses to chemotherapy. Here we compared the MCF-7 breast cancer cells responses to cisplatin in traditional two-dimensional (2D) and in 3D-reconstituted basement membrane (3D-rBM) cell culture models. The results showed a substantial increase of cisplatin resistance mediated by 3D microenvironment. This phenotype was independent of p53 status and autophagy activity and was also observed for other cellular models, including lung cancer cells. Such strong decrease on cellular sensitivity was not due to differences on drug-induced DNA damage, since similar levels of gamma-H2AX and cisplatin-DNA adducts were detected under both conditions. However, the processing of these cisplatin-induced DNA lesions was very different in 2D and 3D cultures. Unlike cells in monolayer, cisplatin-induced DNA damage is persistent in 3D-cultured cells, which, consequently, led to high senescence induction. Moreover, only 3D-cultured cells were able to progress through S cell cycle phase, with unaffected replication fork progression, due to the upregulation of translesion (TLS) DNA polymerase expression and activation of the ATR-Chk1 pathway. Co-treatment with VE-821, a pharmacological inhibitor of ATR, blocked the 3D-mediated changes on cisplatin response, including low sensitivity and high TLS capacity. In addition, ATR inhibition also reverted induction of REV3L by cisplatin treatment. By using REV3L-deficient cells, we showed that this TLS DNA polymerase is essential for the cisplatin sensitization effect mediated by VE-821. Altogether, our results demonstrate that 3D-cell architecture-associated resistance to cisplatin is due to an efficient induction of REV3L and TLS, dependent of ATR. Thus co-treatment with ATR inhibitors might be a promising strategy for enhancement of cisplatin treatment efficiency in breast cancer patients. (AU)

• FAPESP's process: 14/15982-6 - Consequences of repair deficiencies in damaged genome
• Grantee: Carlos Frederico Martins Menck
• Support Opportunities: Research Projects - Thematic Grants
• FAPESP's process: 14/25832-1 - Hippo-YAP pathway as a mediator of extracellular matrix signals in non-malignant and malignant epithelial cells of the mammary gland
• Grantee: Ana Paula Zen Petisco Fiore
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• FAPESP's process: 14/10492-0 - Hippo-YAP as a converging pathway for biochemical and mechanical signals from the extracellular matrix during mammary gland morphogenesis and breast cancer progression
• Grantee: Alexandre Bruni Cardoso
• Support Opportunities: Research Grants - Young Investigators Grants
• FAPESP's process: 11/50856-3 - Role of cellular microenvironment in molecular mechanisms of resistance to chemotherapy in a human mammary carcinoma model
• Grantee: Luciana Rodrigues Gomes
• Support Opportunities: Scholarships in Brazil - Post-Doctoral